Standardization of method for determining glycosylated hemoglobin (Hb A1c) by cation exchange high performance liquid chromatography (original) (raw)
Related papers
Comparison of four chromatographic methods used for measurement of glycated hemoglobin
Revista Romana de Medicina de Laborator, 2016
This parameter’s results accuracy has a special importance in the management of diabetic patients since targets for optimal glycemic control are established using HbA1c values. Several error sources can influence the obtained value, some of them can be counteracted (ex. pipetting errors, storage), and others should be taken into consideration at the interpretation of the result (ex. presence of hemoglobin variants). The aim of this study was to compare four chromatographic methods regarding the costs and the influence of certain error sources on the accuracy of the result. Materials and methods: Samples and controls were analyzed using Variant I, Micromat II and In2it (Bio-Rad) systems, and the BIOMIDI reagent kit for HbA1c measurement. Results: Positive correlation could be observed comparing the results obtained using different methods, except the patients presenting elevated HbF. Pipetting errors modify the results up to 5% in case of Variant I, and up to 10% in case of Micromat ...
The Analytical Performances of Four Different Glycated Hemoglobin Methods
Medicinal Chemistry, 2014
The analytical performances of the Sebia capillary electrophoresis (CE), Roche turbidimetric inhibition immunoassay (TINIA), Tosoh G8 cation-exchange high-performance liquid chromatography (HPLC) and Premier boronate affinity chromatography methods were evaluated. Capillary electrophoresis (CE) was accepted as a comparative method.
A comparative evaluation of glycosylated haemoglobin assays: feasibility of references and standards
Diabetologia, 1984
Four assays; high pressure liquid chromatography, colorimetric with thiobarbituric acid, affinity columns, and microcolumn cation exchange were compared for (1) ability to discriminate between samples taken from diabetic and normal subjects; (2) correlation with each other; (3) stability over time at different temperatures; and (4) reproducibility between laboratories. The most discriminatory (10 samples from a diabetic and 10 samples from a normal group) was the microcolumn cation exchange method (t = 5.25; p < 0.001), but all were significantly different atp < 0.005. The intra-assay coefficient of variation was 1%-6%, except for the affinity column method which was 13% in normal subjects. High pressure liquid chromatography was used as a reference and the other assays correlated well (r=0.93-0.99). Storage at -80 ~ -20 ~ 4 ~ and 24 ~ showed marked differences. The thiobarbituric acid method results were stable except for 24 ~ Microcolumn cation exchange was labile under all conditions. Affinity column was stable for up to 15 days, only if samples were stored as whole blood. High pressure liquid chromatography showed an increase in haemoglobin Ala+b and a decrease in the haemoglobin Ale. Haemoglobin Ale was reproducible for 4 days when stored at 4 ~ and up to 11 days when stored at -80 ~ Samples exchanged between centres at 4 ~ and performed within 5 days by high pressure liquid chromatography for haemoglobin A1 and haemoglobin A1~ correlated well (r= 0.98 and 0.99). Samples exchanged between centres after storage (up to 40 days -80 ~ correlated (r = 0.99) by the thiobarbituric acid method. Thus, standards can be prepared for the thiobarbituric acid method and this method with high pressure liquid chromatography could be used to establish references for clinical assays.
Clinical Biochemistry, 2013
Objectives: Available data on biological variation of HbA 1c revealed marked heterogeneity. We therefore investigated and estimated the components of biological variation for HbA 1c in a group of healthy individuals by applying a recommended and strictly designed study protocol using two different assay methods. Design and methods: Each month, samples were derived on the same day, for three months. Four EDTA whole blood samples were collected from each individual (20 women, 9 men; 20-45 years of age) and stored at −80°C until analysis. HbA 1c values were measured by both high performance liquid chromatography (HPLC) (Shimadzu, Prominence, Japan) and boronate affinity chromatography methods (Trinity Biotech, Premier Hb9210, Ireland). All samples were assayed in duplicate in a single batch for each assay method. Estimations were calculated according to the formulas described by Fraser and Harris. Results: The within subject (CV I)-between subject (CV G) biological variations were 1.17% and 5.58%, respectively for HPLC. The calculated CV I and CV G were 2.15% and 4.03%, respectively for boronate affinity chromatography. Reference change value (RCV) for HPLC and boronate affinity chromatography was 5.4% and 10.4% respectively and individuality index of HbA 1c was 0.35 and 0.93 respectively. Conclusions: This study for the first time described the components of biological variation for HbA 1c in healthy individuals by two different assay methods. Obtained findings showed that the difference between CV A values of the methods might considerably affect RCV. These data regarding biological variation of HbA 1c could be useful for a better evaluation of HbA 1c test results in clinical interpretation.
Comparison of glycated hempglobin with HPLC and capillary electrophoresis
International Journal of Research in Medical Sciences
Background: Hemoglobin A1c, also called A1c or glycated hemoglobin, is hemoglobin with glucose attached. The A1c test evaluates the average amount of glucose in the blood over the last 2 to 3 months. The higher the level of glucose in the blood, the more glycated hemoglobin is formed. Once the glucose binds to the hemoglobin, it remains there for the life of the red blood cell – normally about 120 days. The predominant form of glycated hemoglobin is referred to as A1c. Testing of HbA1c levels via capillary electrophoresis is a relatively new but well-validated method that separates A1c and other Hb fractions via charge difference at high voltage using electro-osmotic flow. This method can be useful in patients who possess such variant hemoglobins because it has a longer runtime, leading to better resolution.Methods: We have processed random samples coming to our laboratory for HbA1C analysis on both the analyzers Biorad D 10 (HPLC method) and Sebia Flex piercing (Capillary Electroph...
Glycohemoglobin assays evaluated in a large-scale quality-control survey
Clinical chemistry, 1995
We report the results of a national quality-control survey on glycohemoglobin (GHb), monitored in France by the Société Française de Biologie Clinique on behalf of the authority of the "Agence du Médicament." A sample of lyophilized hemolysate was sent to 3109 laboratories. Results were obtained from 2770 laboratories. HbA1C, HbA1, and total GHb were measured by 50%, 24%, and 26% of the participants, respectively. Of these measurements, 79% of the HbA1C results and 76% of the total GHb results, but only 48% of the HbA1 results, were within the +/- 20% limits of the indicated target values. Mean values for the hemolysate ranged from 8% to 11% for HbA1C, from 7% to 12% for HbA1, and from 11% to 13% for total GHb. The interlaboratory CVs ranged from 3% to 20%, according to method used. So, methods used for GHb assay, which are based on various principles, exhibit very different analytical performances. Nonetheless, this large-scale study indicates that some techniques can sup...